PROJECT 2 ? Summary Goal: Our goal is to determine how tumor-specific replication abnormalities can be used to develop more effective anti-cancer therapies for adult acute myelogenous leukemia (AML) and glioblastoma multiforme (GBM), two rapidly lethal, treatment-refractory tumors. Our organizing hypothesis is that aberrant replication is common in AML and GBM, and represents an opportunity to improve treated survival by identifying drugs, small molecules and treatment regimens that synergize with replication defects to promote cell killing. Innovations: The Project 2 Research Plan has the following innovative features: ? Replication and replication abnormalities as tumor-specific therapeutic vulnerabilities in AML and GBM, a focus viewed as ?very novel? in prior review. ? DNA replication fork dynamics analyzed at the single molecule level. ? Network modeling to link replication abnormalities to genomic instability and therapeutic response. ? Microfluidic culture of GBM tissue on a novel platform to enable multiplexed drug profiling. ? Mixed-genotype competitive growth assays to determine mutant fitness?and thus potential therapeutic importance?in culture and in xenografts. Approach/Aims: We have three inter-related Aims collectively focused on defining DNA replication behavior and therapeutic response in AML, GSCs and GBM tissue. Aim 1: Define AML replication dynamics and the role of aberrant replication in therapeutic response. Aim 2: Define GBM replication dynamics and the role of aberrant replication in therapeutic response. Aim 3: Identify mutation-dependent mechanisms that drive subclonal heterogeneity and therapeutic resistance (a collaborative Project Aim). Impact: The proposed research will provide or develop: ? Detailed picture of mutation-dependent replication dynamics in AML and GBM. ? Link altered replication dynamics influence therapeutic response in AML and GBM. ? Identify or predict potentially effective new therapies and regimens for AML and GBM. ? Test potential new therapeutic vulnerabilities using clinically relevant preclinical models.